pollen morphology of philippine species of phyllanthus

copy 2009 Nationaal Herbarium Nederland

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Blumea 54 2009 47ndash58wwwingentaconnectcomcontentnhnblumea doi103767000651909X474087RESEARCH ARTICLE

INTRODUCTION

Phyllanthus is the largest genus of the Phyllanthaceae (Eushyphorbiaceae sl) It contains about 830 species (Govaert et al 2000) On the basis of a large molecular analysis of Phyllanthus and its relatives Kathriarachchi et al (2006) recommended the inclusion of Breynia Glochidion Reverchonia and Sauropus to form a large monophyletic genus Phyllanthus sl This genus would then comprise c 1 269 species (Govaert et al 2000) They found that the subgenera Conami Emblica Eriococcus Gomphidium (sensu Schmid 1991) and Xylophylla (except for P sellowianus) are monophyletic whereas the subgenera Isocladus Kirganelia and Phyllanthus appeared to be not monophyletic Most clades obtained in their analyses were supported by palynological characters

In the study of the West Indian species of Phyllanthus Web-ster (1956) discovered the extraordinary diversity of pollen morphology within the species using light microscope (LM) Punt (1967) showed two series of Phyllanthus pollen types and proposed their evolutionary trends The precise usage of pollen characters in taxonomy for the small pollen size usually around 20ndash30 μm diam is difficult therefore the aid of scanning electronic microscope (SEM) in the observation of the exine ornamentation is necessary At least 30 pollen types were reported in 220 species of Phyllanthus studied using LM andor SEM (Erdtman 1952 Webster 1956 1986 Koumlhler 1965 1967 Punt 1967 1972 1980 1987 Punt amp Rentrop 1973 Bor 1979 Meewis amp Punt 1983 Long amp Yu 1984 Rossignol et al 1987 Lobreau-Callen et al 1988 Chen amp Wu 1997 Webster amp Carpenter 2002 Wei et al 2002 Sagun amp Van der Ham 2003 Santiago et al 2004)

In the Philippines more than 30 species of Phyllanthus have been reported previously and can be classified into five sub-genera and eleven sections (Elmer 1908 1910 1911 Robin-son 1909 1911 Merrill 1912 1914 1920 1923 Quisumbing amp Merrill 1928 Airy Shaw 1983 Hoffmann et al 2003) The

minute unisexual flowers and the variable leaf morphology caused the classification of some species to remain in doubt In order to provide new evidence for the species classification we undertook the present pollen morphological study of Philip-pine Phyllanthus species

MATERIAL AND METHODS

Pollen samples were collected from specimens deposited in the following herbaria L NHU PUH (Table 1)

Per sample anthers of 1ndash3 flowers were placed in 15 ml centri-fuge tubes immersed in acetic acid for 30 min until soft crushed by a glass bar to release the pollen grains and then acetolyzed (Erdtman 1952) The acetolyzed pollen was then dehydrated in a series of 70 90 and 100 ethanol and pipeted onto a filter membrane to be air dried Under a thin layer of gold coating the pollen grains were examined and photographed using a Hitachi S-800 Scanning Electron Microscope Pollen size was based on measuring 2ndash4 pollen grains per sample The terminology used in this paper follows Huang (1972) and Punt et al (2007)

RESULTS

Eleven pollen types were recognized in the Philippine Phylshylanthus species studied (Table 2) seven of which have been reported previously and four (P erythrotrichus type P gloshychidioides type P securinegoides type Pilate type) might be reported here for the first time A key to the pollen types is presented below

Key to the pollen types

1 Pollen grain pantoporate P buxifolius type1 Pollen grain colporate syncolporate or areolate (ie panto-

syncolpate) 2

2 Pollen grain syncolporate or areolate (ie pantosyncol-pate) 3

2 Pollen grain colporate 5

3 Pollen grain areolate (ie pantosyncolpate) with 16ndash26 colpi P virgatus type

3 Pollen grain syncolporate with three colpi 4

Pollen morphology of Philippine species of Phyllanthus (Phyllanthaceae Euphorbiaceae sl)

Y-J Chen1 S-H Chen1 T-C Huang2 M-J Wu1

1 Graduate Institute of Biological Resources and Technology National Dong Hwa University Meilun Campus formerly known as National Hualien Univer-sity of Education Hualien 970 Taiwan

2 Institute of Plant Biology National Taiwan University Taipei 106 Taiwan

Abstract The pollen morphology of 21 Philippine Phyllanthus species belonging to five subgenera and eleven sections was studied using scanning electron microscopy Eleven pollen types were recognized of which seven were previously reported and four are newly described ie the Phyllanthus erythrotrichus type the P glochidioides type the P securinegoides type and the Pilate type The pollen morphology of the Philippine Phyllanthus species included in this study provides insight into the taxonomy as well as the phylogeny of these species

Published on 30 October 2009

Key words

EuphorbiaceaePhilippinesPhyllanthaceaePhyllanthuspollen morphologySEMtaxonomy

48 Blumea ndash Volume 54 2009

Species Location Voucher

P acidus (L) Skeels Santa Luzon Gaerlan FJM et al PPI 10249 (L) P amarus Schumach amp Thonn Palawan Wu M-J 2951 (NHU) P benguetensis CBRob Luzon Merrill ED 9684 (L) P buxifolius (Blume) MuumlllArg Bohol Ramos M BS 43248 (L) P erythrotrichus CBRob Susong-Dalaga Luzon Robinson CB BS 29333 (L) P glochidioides Elmer Paraiso Luzon Edantildeo GE PNH 40177 (L) P kinabaluicus Airy Shaw Beaufort Mountain Palawan Ridsdale CE SMHI 241 (L) P lamprophyllus MuumlllArg Palawan Gaerlan FJM PPI 13214 (L) P lanceifolius Merr Catanduanes Ramos M amp G Edano BS 75281 (PUH) P megalanthus CBRob Luzon Curran HM FB 17243 (L) P mindorensis CBRob Lobo Luzon Sulit MD PNH 15718 (L) P ramosii Quisumb amp Merr Luzon Mabesa C 853 (L) P reticulatus Poir Los Bantildeos Luzon Hallier JG 4117 (L) P samarensis MuumlllArg Paranas Samar Reynoso EJ PPI 7550 (L) P securinegoides Merr Panacan Palawan Sulit MD PNH 16268 (L) P sibuyanensis Elmer Magallanes Sibuyan Elmer ADE 12113 (L) P urinaria ssp nudicarpus Rossignol amp Haicour Palawan Wu M-J 2952 (NHU) P urinaria ssp urinaria L Palawan Wu M-J 2954 (NHU) P virgatus GForst Palawan Wu M-J 2953 (NHU) P aff glochidioides Tagkawayan Luzon Lagrimas MQ PNH 39954 (L) P aff samarensis Santa Cruz Luzon Ridsdale CE1479 (L) Phyllanthus sp A Bohol Ramos M BS 43203 (L)

Table1 Locality and voucher specimens of the Philippine Phyllanthus species sampled in this study

4 Lumina of the reticulum with granules P maritimus type4 Lumina of the reticulum without granules P casticum type

5 Pollen grain 3-colporate 65 Pollen grain 4- or 5-colporate 8

6 Colpi nearly confluent at the poles exine ornamentation rugulate-reticulate P acidus type

6 Colpi not confluent at the poles exine ornamentation bi-reticulate or striate-reticulate 7

7 Exine ornamentation bi-reticulate pollen grain subprolate to prolate P amarus type

7 Exine ornamentation striate-reticulate pollen grain prolate P erythrotrichus type

8 Exine ornamentation pilate colpi obscure Pilate type 8 Exine ornamentation reticulate or bi-reticulate colpi dis-

tinct 9

9 Exine ornamentation bi-reticulate pollen grain 4- or 5-col-porate P urinaria type

9 Exine ornamentation reticulate pollen grain 4-colporate 10

10 Colpus margins smooth muri round verrucate P securinegoides type

10 Colpus margins ragged muri coniculate P glochidioides type

Fig 1 Scanning electron micrographs of the Phyllanthus erythrotrichus type andashf P erythrotrichus mdash Scale bars andashc = 5 microm dndashf = 2 microm

cb

d

a

e f

49Y-J Chen et al Pollen morphology of Phyllanthus

DESCRIPTION OF THE POLLEN TyPES

I Phyllanthus erythrotrichus type mdash Fig 1

Pollen grains prolate to perprolate P = 153ndash193 microm E = 93ndash100 microm PE = 158ndash208 Aperture system mdash 3-colporate Ornamentation mdash Striate-reticulate lumina small and nar-row Species included mdash P erythrotrichus Note mdash The ornamentation is similar to that of the P urinaria type but the lumina are narrower

II Phyllanthus virgatus type mdash Fig 2 3

Pollen grains spheroidal to ellipsoidal Polarity unknown Size = 153ndash225 by 150ndash200 microm Aperture system mdash Areolate (ie pantosyncolpate) Ornamentation mdash Reticulate to coarsely reticulate Species included mdash P lanceifolius P samarensis P virgashytus P aff samarensis Note mdash The number of lumina per areola varies P lanceifoshylius and P samarensis plusmn 30 P virgatus plusmn 60 and Phyllanthus aff samarensis plusmn 70 Phyllanthus aff samarensis differs from the other species of the P virgatus type by its colpus margins consisting of two parallel muri rather than four References mdash Punt 1980 1987 Long amp Yu 1984 Webster 1986 Lobreau-Callen et al1988 Chen amp Wu 1997

III Phyllanthus casticum type mdash Fig 4

Pollen grains oblate spheroidal P = 111ndash117 microm E = 117ndash125 microm PE = 091ndash096 Aperture system mdash 3-syncolporate Ornamentation mdash Reticulate Species included mdash P reticulatus Note mdash This type is similar to the P acidus type but differs by its confluent colpi References mdash Koumlhler 1965 Punt 1967 1980 Bor 1979 Long amp Yu 1984 Lobreau-Callen et al 1988

IV Phyllanthus acidus type mdash Fig 5

Pollen grains oblate spheroidal P = 123ndash133 microm E = 133ndash 150 microm PE = 089ndash095 Aperture system mdash 3-colporate colpi nearly confluent at the poles Ornamentation mdash Rugulate-reticulate Species included mdash P acidus References mdash Koumlhler 1965 1967 Punt 1967

V Phyllanthus amarus type

Pollen grains prolate spheroidal to subprolate P = 153ndash170 microm E = 130ndash157 microm PE = 106ndash131 Aperture system mdash 3-colporate Ornamentation mdash Bi-reticulate Species included mdash P amarus Note mdash This type and the P urinaria type have both bi- reticulate ornamentation but they differ in the number of colpi References mdash Punt amp Rentrop 1973 Bor 1979 Punt 1980 Long amp Yu 1984 Chen amp Wu 1997

Sub

genu

s S

ectio

n P

olle

n ty

pe

Taxa

A

pert

ure

Equ

ator

ial s

hape

S

ize

P b

y E

(micro

m)

PE

O

rnam

enta

tion

Erio

cocc

us

Sce

pasm

a P

bux

ifoliu

s ty

pe

P b

uxifo

lius

num

erou

s sp

hero

idal

to e

llips

oida

l 22

2ndash

224

by

244

ndash25

6

ndash re

ticul

ate

E

mbl

icas

trum

P

bux

ifoliu

s ty

pe

P la

mpr

ophy

llus

num

erou

s sp

hero

idal

to e

llips

oida

l 22

9ndash

246

by

213

ndash22

9

ndash re

ticul

ate

E

rioco

ccus

P

bux

ifoliu

s ty

pe

P k

inab

alui

cus

num

erou

s sp

hero

idal

to e

llips

oida

l 20

0ndash

204

by

200

ndash20

4

ndash re

ticul

ate

P

meg

alan

thus

nu

mer

ous

sphe

roid

al to

elli

psoi

dal

250

by

244

ndash26

1

ndash re

ticul

ate

P

min

dore

nsis

nu

mer

ous

sphe

roid

al to

elli

psoi

dal

175

ndash18

0 by

18

0 ndash

retic

ulat

e

P s

ibuy

anen

sis

num

erou

s sp

hero

idal

to e

llips

oida

l 18

0ndash

200

by

183

ndash19

6 ndash

retic

ulat

e

Gom

phid

ium

A

deno

gloc

hidi

on

P m

ariti

mus

type

P

ram

osii

3-sy

ncol

pora

te

obla

te s

pher

oida

l 15

8ndash1

67

by 1

71

092

ndash0

98

retic

ulat

e

Gom

phid

ium

P

glo

chid

ioid

es ty

pe

P g

loch

idio

ides

4-

colp

orat

e su

bpro

late

21

7ndash

221

by

175

ndash18

8 1

15ndash1

26

retic

ulat

e

P a

ff g

loch

idio

ides

4-

colp

orat

e su

bpro

late

19

1ndash

208

by

158

ndash17

5 1

19ndash1

21

retic

ulat

e

P

sec

urin

egoi

des

type

P

sec

urin

egoi

des

4-co

lpro

late

pr

olat

e sp

hero

idal

22

9ndash

243

by

219

ndash22

9

100

ndash11

1 re

ticul

ate

Isoc

ladu

s P

arap

hylla

nthu

s P

ery

thro

tric

hus

type

P

ery

thro

tric

hus

3-co

lpor

ate

prol

ate

to p

erpr

olat

e 15

3ndash1

93

by 9

3ndash1

00

158

ndash2

08

stria

ted

retic

ulat

e

Par

aphy

llant

hus

P v

irgat

us ty

pe

P la

ncei

foliu

s ar

eola

te

sphe

roid

al to

elli

psoi

dal

175

ndash22

5 b

y 17

5ndash1

96

ndash co

arse

ly r

etic

ulat

e

Mac

raea

P

virg

atus

type

P

sam

aren

sis

areo

late

sp

hero

idal

to e

llips

oida

l 15

3ndash1

67

by 1

50

ndash16

3 ndash

coar

sely

ret

icul

ate

P

virg

atus

ar

eola

te

sphe

roid

al to

elli

psoi

dal

167

ndash18

8 by

15

8ndash1

88

ndash re

ticul

ate

P

aff

sam

aren

sis

ar

eola

te

sphe

roid

al to

elli

psoi

dal

183

ndash19

6 by

19

2ndash

200

ndash

retic

ulat

e

Kirg

anel

ia

Ani

sone

ma

P c

astic

um ty

pe

P r

etic

ulat

us

3-sy

ncol

pora

te

obla

te s

pher

oida

l 11

1ndash1

17

by 1

17ndash

125

0

91ndash

096

re

ticul

ate

C

icca

P

aci

dus

type

P

aci

dus

3-co

lpor

ate

obla

te s

pher

oida

l 12

3ndash1

33

by 1

33

ndash15

0 0

89ndash

095

ru

gula

te r

etic

ulat

e

Phy

llant

hus

Phy

llant

hus

P a

mar

us ty

pe

P a

mar

us

3-co

lpor

ate

prol

ate

sper

oida

l to

subp

rola

te

153

ndash17

0 by

13

0ndash1

57

106

ndash13

1 bi

retic

ulat

e

Urin

aria

P

urin

aria

type

P

ben

guet

ensi

s 4-

colp

rola

te

prol

ate

200

ndash22

1 b

y 12

1ndash1

33

163

ndash17

6 bi

retic

ulat

e

P u

rinar

ia s

sp n

udic

arpu

s 4-

colp

rola

te

prol

ate

225

ndash23

8 b

y 12

9ndash1

46

163

ndash17

4 bi

retic

ulat

e

P u

rinar

ia s

sp u

rinar

ia

4-co

lpro

late

pr

olat

e 16

0ndash1

63

by 1

1ndash12

1

36ndash1

45

bire

ticul

ate

not a

ssig

ned

not a

ssig

ned

Pila

te ty

pe

Phy

llant

hus

sp A

4-

colp

orat

e su

bpro

late

14

3 b

y 11

ndash12

3 1

16ndash1

30

pila

te

Tab

le 2

P

olle

n ty

pes

of P

hilip

pine

Phy

llant

hus

spec

ies


Fig 2 Scanning electron micrographs of the Phyllanthus virgatus type andashf P lanceifolius gndashl P samarensis mdash Scale bars andashc gndashi = 5 microm dndashf jndash l = 2 microm

cb

d

a

g h i

j k l

e f


Fig 3 Scanning electron micrographs of the Phyllanthus virgatus type andashf P virgatus gndashl P aff samarensis mdash Scale bars andashc gndashi = 5 microm dndashf jndash l = 2 microm

cb

d

a

g h i

j k l

e f


Fig 4 Scanning electron micrographs of the Phyllanthus casticum type andashf P reticulatus mdash Scale bars andashc = 5 microm dndashf = 2 microm

cb

d

a

e f

Fig 5 Scanning electron micrographs of the Phyllanthus acidus type andashf P acidus mdash Scale bars andashc = 5 microm dndashf = 2 microm

cb

d

a

e f


VI Phyllanthus urinaria type mdash Fig 6

Pollen grains prolate P = 160ndash238 microm E = 1105ndash146 microm PE = 136ndash176 Aperture system mdash 4- or 5-colporate colpi narrow Ornamentation mdash Bi-reticulate Species included mdash P benguetensis P urinaria (both sub-species) Note mdash Rossignol et al (1987) found that species in sect Urinaria with chromosome number 2n = 50 have 4-colporate pollen while species with chromosome number 2n = 100 have 5-colporate pollen References mdash Punt amp Rentrop 1973 Bor 1979 Long amp Yu 1984 Rossignol et al 1987 Chen amp Wu 1997

VII Phyllanthus buxifolius type mdash Fig 7

Pollen grains spheroidal to ellipsoidal Polarity unknown Size = 175ndash253 by 180ndash261 microm Aperture system mdash Pantoporate Ornamentation mdash Coarsely reticulate less than 10 granules per lumen Species included mdash P buxifolius P kinabaluicus P lamshyprophyllus P megalanthus P mindorensis P sibuyanensis References mdash Punt 1980 1987 Long amp Yu 1984 Wei et al 2002

VIII Phyllanthus maritimus type mdash Fig 8

Pollen grains oblate spheroidal P = 158ndash167 microm E = 171 microm PE = 092ndash098 Aperture system mdash 3-syncolporate Ornamentation mdash Reticulate lumina with small granules Species included mdash P ramosii

Note mdash The P maritimus type is similar to the P casticum type but differs by the presence of granules in the lumina References mdash Punt 1980 Meewis amp Punt 1983

IX Phyllanthus glochidioides type mdash Fig 9

Pollen grains subprolate P = 171ndash221 microm E = 158ndash188 microm PE = 115ndash126 Aperture system mdash 4-colporate colpi slit-like with ragged margins Ornamentation mdash Coarsely reticulate Species included mdash P glochidioides Phyllanthus aff glochishydioides

X Phyllanthus securinegoides type mdash Fig 10

Pollen grains prolate spheroidal P = 229ndash243 microm E = 219ndash 229 microm PE = 100ndash111 Aperture system mdash 4-colporate colpi with smooth margins Ornamentation mdash Reticulate Species included mdash P securinegoides

XI Pilate type mdash Fig 11

Pollen grains subprolate P = 143 microm E = 110ndash123 microm PE = 116ndash130 Aperture system mdash 4-colporate colpi obscure using SEM Ornamentation mdash Pilate pila mostly with rounded capita Note mdash This type is similar to the inaperturate pollen grains of P attenuatus (subg Conami sect Hylaeanthus Webster amp Carpenter 2002) but differs by the presence of colpi Species included mdash Phyllanthus sp A

Fig 6 Scanning electron micrographs of the Phyllanthus urinaria type andashf P benguetensis mdash Scale bars andashc = 5 microm dndashf = 2 microm

cb

d

a

e f


Fig 7 Scanning electron micrographs of the Phyllanthus buxifolius type a d P buxifolius b e P kinabaluicous c f P lamprophyllus g j P megalanthus h k P mindorensis i l P sibuyanensis mdash Scale bars andashc gndashi = 5 microm dndashf jndash l = 2 microm

cb

d

a

g h i

j k l

e f


DISCUSSION

Pollen types and infrageneric classification

Five of the 11 pollen types in Philippine Phyllanthus each occur in a single section (Table 2) The P amarus type and P urinaria type both belonging to subg Phyllanthus have the same bi-reticulate ornamentation but differ in the number of colpi The P acidus type P casticum type and P maritimus type share reticulate exine ornamentation However the P maritimus type has granules in the lumina The P acidus type and P casticum type (both subg Kirganelia) can be distinguished by the length of their colpi

Two pollen types occur in more than one section the P virgatus type in the two sections of subg Isocladus the P buxifolius type in the three sections of subg Eriococcus The P buxifolius type found in six species is homogeneous The P virgatus type found in four species varies in the morphology of the colpus margins Phyllanthus aff samarensis has colpus margins con-sisting of a single murus while the other species have colpus margins comprising two muri Therefore it might be appropriate to describe a different subtype for P aff samarensis Evidently the areolate pollen of P lanceifolius (sect Paraphyllanthus Merrill 1914) is closer to that of sect Macraea than to the 3-colporate pollen of P erythrotrichus (sect Paraphyllanthus Punt 1967 1987) Transferring P lanceifolius to sect Macraea should therefore be considered

Pollen morphologically subg Gomphidium including three pol-len types is most heterogeneous in Philippine Phyllanthus The P maritimus type (sect Adenoglochidion) is 3-syncolporate The P glochidioides type and P securinegoides type (both sect Gomphidium) have 4-colporate pollen They differ by their exine ornamentation and the morphology of the colpus margins

Merrill (1914) found P securinegoides hard to classify He noticed that P securinegoides did not fit in sect Gomphidium because the scarce styles are united spreading and promi-nently cleft Hence he suggested that a new section should be described Pollen morphology confirms Merrillrsquos opinion

Phyllanthus sp A has not been assigned to a section and sub-genus yet Its pollen (Pilate type) is similar to the pilate pollen of P attennutus (subg Conami sect Hylaeanthus Webster amp Carpenter 2002) but has a 4-colporate apertural system

Pollen types and phylogenetic relationships

Subgenera Isocladus Kirganelia and Phyllanthus are not monophyletic based on ITS and matK DNA sequence analy-ses (Kathriarachchi et al 2006) According to these data sect Paraphyllanthus is more primitive than sect Macraea (subg Isocladus) sect Anisonema more primitive than sect Cicca (subg Kirganelia) and P amarus more primitive than sect Urishynaria (subg Phyllanthus) Punt (1967) considered an increased number of colpi as a derived feature Thus the P erythrotrichus type of sect Paraphyllanthus would be more primitive than the P virgatus type of sect Macraea and the P amarus type of sect Phyllanthus is more primitive than the P urinaria type of sect Urinaria In these cases the pollen morphological trend proposed by Punt is concordant with the DNA sequence data According to Punt (1967) the colporate P acidus type of sect Cicca is more primitive than the syncolporate P casticum type of sect Anisonema This conflicts with the DNA sequence data However comparing the ornamentation in detail the P casticum type of sect Anisonema has a primitive reticulate ornamentation and the P acidus type of sect Cicca has an advanced rugulate-reticulate ornamentation This result tells us that the ornamentation trend is concordant with the DNA sequence data of Kathriarachchi et al (2006)

Fig 8 Scanning electron micrographs of the Phyllanthus maritimus type andashf P ramosii mdash Scale bars andashc = 5 microm dndashf = 2 microm

cb

d

a

e f


Fig 9 Scanning electron micrographs of the Phyllanthus glochidioides type andashf P glochidioides gndashl Phyllanthus aff glochidioides mdash Scale bars andashc gndashi = 5 microm dndashf jndash l = 2 microm

cb

d

a

g h i

j k l

e f


Fig 11 Scanning electron micrographs of the Pilate type andashf Phyllanthus sp A mdash Scale bars andashc = 5 microm dndashf = 2 microm

Fig 10 Scanning electron micrographs of the Phyllanthus securinegoides type andashf P securinegoides mdash Scale bars andashc = 5 microm dndashf = 2 microm

cb

d

a

e f

cb

d

a

e f


Acknowledgements We would like to express our sincere thanks to the directors of the National Herbarium Nederland (L) and the Jose Vera Santos Memorial Herbarium (PUH) for the loan of specimens and providing pollen materials We appreciate the anonymous reviewer for providing advice and editing our English and Dr Peter van Welzen and Mr Leonardo Co for providing references Thanks also to Ms Jin-Yen Lin and Ms Ya-Yun Yang National Taiwan University for their assistance in using the scanning electron microscope This study was supported in part by the National Science Council (grant no NSC95-2621-B-026-001) Executive Yuan Taiwan

REFERENCES

Airy Shaw HK 1983 An alphabetical enumeration of the Euphorbiaceae of the Philippine Islands Royal Botanical Gardens Kew

Bor J 1979 Pollen morphology and the bi-reticulate exine of the Phyllanthus species (Euphorbiaceae) from Mauritius and Reunion Review of Paleo-botany and Palynology 27 149ndash172

Chen S-H Wu M-J 1997 A revision of the herbaceous Phyllantus L (Eu-phorbiaceae) in Taiwan Taiwania 42 239ndash261

Elmer ADE 1908 A century of new plants Leaflets of Philippine Botany 1 272ndash359

Elmer ADE 1910 Euphorbiaceae collected on Sibuyan Island Leaflets of Philippine Botany 3 901ndash931

Elmer ADE 1911 Euphorbiaceae collected on Palawan Island Leaflets of Philippine Botany 4 1271ndash1306

Erdtman G 1952 Pollen morphology and plant taxonomy angiosperms Almquist amp Wiksell Stockholm

Govaert R Frodin DG Radcliffe-Smith A 2000 World checklist and biblio-graphy of Euphorbiaceae vol 4 Royal Botanic Gardens Kew

Hoffmann P Barker AJM Madulid DA Proctor J 2003 Phyllanthus balgooyi (Euphorbiaceae sl) a new nickel-hyperaccumulating species from Pala-wan and Sabah Blumea 48 193ndash199

Huang T-C 1972 Pollen flora of Taiwan 1st ed Botany Department National Taiwan University Taipei 106 Taiwan

Kathriarachchi H Samuel R Hoffmann P Mlinarec J Wurdack KJ Ralima-nana H Stuessy TF Chase MW 2006 Phylogenetics of tribe Phyllantheae (Phyllanthaceae Euphorbiaceae sensu lato) based on nrITS and plastid matK DNA sequence data American Journal of Botany 93 637ndash655

Koumlhler E 1965 Die Pollenmorphologie der biovulaten Euphorbiaceae und ihre Bedeutung fuumlr die Taxonomie Grana Palynologica 6 26ndash120

Koumlhler E 1967 Uumlber Beziehungen zwischen Pollenmorphologie und Poly- ploidiestufen im Verwandtschaftsbereich der Gattung Phyllanthus (Euphor-biaceae) Feddes Repertorium 74 159ndash165

Lobreau-Callen D Punt W Schmid M 1988 Pollen morphology and taxon-omy of the Phyllanthus species (Euphorbiaceae) native to New Caledonia Review of Paleobotany and Palynology 53 283ndash304

Long H Yu C-H 1984 Intrageneric variation of pollen types in the genus Phyllanthus L Acta Botanica Sinica 26 247ndash251

Meewis B Punt W 1983 Pollen morphology and taxonomy of the subgenus Kirganelia (Jussieu) Webster (genus Phyllanthus Euphorbiaceae) from

Africa Review of Paleobotany and Palynology 39 131ndash160Merrill ED 1912 Notes on Philippine Euphorbiaceae Philippine Journal of

Science Section C Botany 7 379ndash410Merrill ED 1914 Notes on Philippine Euphorbiaceae II Philippine Journal

of Science Section C Botany 9 461ndash493Merrill ED 1920 Notes on Philippine Euphorbiaceae III Philippine Journal

of Science 16 539ndash579Merrill ED 1923 An enumeration of Philippine flowering plants 2 391ndash396

Bureau of Printing ManilaPunt W 1967 Pollen morphology of the genus Phyllanthus (Euphorbiaceae)

Review of Paleobotany and Palynology 3 141ndash150Punt W 1972 Pollen morphology and taxonomy of section Ceramanthus

Baillon sl of the genus Phyllanthus (Euphorbiaceae) Review of Paleo-botany and Palynology 13 213ndash228

Punt W 1980 Pollen morphology of the Phyllanthus species (Euphorbia-ceae) occurring in New Guinea Review of Paleobotany and Palynology 31 155ndash177

Punt W 1987 A survey of pollen morphology in Euphorbiaceae with special reference to Phyllanthus Botanical Journal of the Linnean Society 94 127ndash142

Punt W Hoen PP Blackmore S Nilsson S Le Thomas A 2007 Glossary of pollen and spore terminology Review of Paleobotany and Palynology 143 1ndash81

Punt W Rentrop J 1973 Pollen morphology of the Phyllanthus species (Euphorbiaceae) occurring in the continental United States Review of Paleobotany and Palynology 16 243ndash261

Quisumbing E Merrill ED 1928 New Philippine plants Philippine Journal of Science 37 133ndash212

Robinson CB 1909 Philippine Phyllanthinae Philippine Journal of Science Section C Botany 4 71ndash106

Robinson CB 1911 Alabastra Philippinensia III Philippine Journal of Sci-ence Section C Botany 6 319ndash358

Rossignol L Rossignol M Haicour R 1987 A systematic revision of Phyl-lanthus subsection Urinaria (Euphorbiaceae) American Journal of Botany 74 1853ndash1862

Sagun VG Van der Ham RWJM 2003 Pollen morphology of the Flueggeinae (Euphorbiaceae Phyllanthoideae) Grana 42 193ndash219

Santiago LJM Louro RP Emmerich M Barth OM 2004 The pollen mor-phology of Phyllanthus (Euphorbiaceae) section Choretropsis Botanical Journal of the Linnean Society 144 243ndash250

Schmid M 1991 Phyllanthus In Morat P Mackee HS (eds) Flore de la Nouvelle-Caleacutedonie et Deacutependances 17 31ndash320 Museacuteum National drsquoHistoire Naturelle Paris France

Webster GL 1956 A monographic study of the West Indian species of Phyl-lanthus Journal of the Arnold Arboretum 37 217ndash268

Webster GL 1986 A revision of Phyllanthus (Euphorbiaceae) in Eastern Melanesia Pacific Science 40 88ndash105

Webster GL Carpenter K 2002 Pollen morphology and phylogenetic rela-tionships in neotropical Phyllanthus (Euphorbiaceae) Botanical Journal of the Linnean Society 138 325ndash338

Wei W-Z Kiu H-S Zhou L-H 2002 Pollen morphology of Phyllanthus (Eu-phorbiaceae) Acta Botanica Yunnanica 24 253ndash259


Species Location Voucher

P acidus (L) Skeels Santa Luzon Gaerlan FJM et al PPI 10249 (L) P amarus Schumach amp Thonn Palawan Wu M-J 2951 (NHU) P benguetensis CBRob Luzon Merrill ED 9684 (L) P buxifolius (Blume) MuumlllArg Bohol Ramos M BS 43248 (L) P erythrotrichus CBRob Susong-Dalaga Luzon Robinson CB BS 29333 (L) P glochidioides Elmer Paraiso Luzon Edantildeo GE PNH 40177 (L) P kinabaluicus Airy Shaw Beaufort Mountain Palawan Ridsdale CE SMHI 241 (L) P lamprophyllus MuumlllArg Palawan Gaerlan FJM PPI 13214 (L) P lanceifolius Merr Catanduanes Ramos M amp G Edano BS 75281 (PUH) P megalanthus CBRob Luzon Curran HM FB 17243 (L) P mindorensis CBRob Lobo Luzon Sulit MD PNH 15718 (L) P ramosii Quisumb amp Merr Luzon Mabesa C 853 (L) P reticulatus Poir Los Bantildeos Luzon Hallier JG 4117 (L) P samarensis MuumlllArg Paranas Samar Reynoso EJ PPI 7550 (L) P securinegoides Merr Panacan Palawan Sulit MD PNH 16268 (L) P sibuyanensis Elmer Magallanes Sibuyan Elmer ADE 12113 (L) P urinaria ssp nudicarpus Rossignol amp Haicour Palawan Wu M-J 2952 (NHU) P urinaria ssp urinaria L Palawan Wu M-J 2954 (NHU) P virgatus GForst Palawan Wu M-J 2953 (NHU) P aff glochidioides Tagkawayan Luzon Lagrimas MQ PNH 39954 (L) P aff samarensis Santa Cruz Luzon Ridsdale CE1479 (L) Phyllanthus sp A Bohol Ramos M BS 43203 (L)

Table1 Locality and voucher specimens of the Philippine Phyllanthus species sampled in this study

4 Lumina of the reticulum with granules P maritimus type4 Lumina of the reticulum without granules P casticum type

5 Pollen grain 3-colporate 65 Pollen grain 4- or 5-colporate 8

6 Colpi nearly confluent at the poles exine ornamentation rugulate-reticulate P acidus type

6 Colpi not confluent at the poles exine ornamentation bi-reticulate or striate-reticulate 7

7 Exine ornamentation bi-reticulate pollen grain subprolate to prolate P amarus type

7 Exine ornamentation striate-reticulate pollen grain prolate P erythrotrichus type

8 Exine ornamentation pilate colpi obscure Pilate type 8 Exine ornamentation reticulate or bi-reticulate colpi dis-

tinct 9

9 Exine ornamentation bi-reticulate pollen grain 4- or 5-col-porate P urinaria type

9 Exine ornamentation reticulate pollen grain 4-colporate 10

10 Colpus margins smooth muri round verrucate P securinegoides type

10 Colpus margins ragged muri coniculate P glochidioides type

Fig 1 Scanning electron micrographs of the Phyllanthus erythrotrichus type andashf P erythrotrichus mdash Scale bars andashc = 5 microm dndashf = 2 microm

cb

d

a

e f













Sub

genu

s S

ectio

n P

olle

n ty

pe

Taxa

A

pert

ure

Equ

ator

ial s

hape

S

ize

P b

y E

(micro

m)

PE

O

rnam

enta

tion

Erio

cocc

us

Sce

pasm

a P

bux

ifoliu

s ty

pe

P b

uxifo

lius

num

erou

s sp

hero

idal

to e

llips

oida

l 22

2ndash

224

by

244

ndash25

6

ndash re

ticul

ate

E

mbl

icas

trum

P

bux

ifoliu

s ty

pe

P la

mpr

ophy

llus

num

erou

s sp

hero

idal

to e

llips

oida

l 22

9ndash

246

by

213

ndash22

9

ndash re

ticul

ate

E

rioco

ccus

P

bux

ifoliu

s ty

pe

P k

inab

alui

cus

num

erou

s sp

hero

idal

to e

llips

oida

l 20

0ndash

204

by

200

ndash20

4

ndash re

ticul

ate

P

meg

alan

thus

nu

mer

ous

sphe

roid

al to

elli

psoi

dal

250

by

244

ndash26

1

ndash re

ticul

ate

P

min

dore

nsis

nu

mer

ous

sphe

roid

al to

elli

psoi

dal

175

ndash18

0 by

18

0 ndash

retic

ulat

e

P s

ibuy

anen

sis

num

erou

s sp

hero

idal

to e

llips

oida

l 18

0ndash

200

by

183

ndash19

6 ndash

retic

ulat

e

Gom

phid

ium

A

deno

gloc

hidi

on

P m

ariti

mus

type

P

ram

osii

3-sy

ncol

pora

te

obla

te s

pher

oida

l 15

8ndash1

67

by 1

71

092

ndash0

98

retic

ulat

e

Gom

phid

ium

P

glo

chid

ioid

es ty

pe

P g

loch

idio

ides

4-

colp

orat

e su

bpro

late

21

7ndash

221

by

175

ndash18

8 1

15ndash1

26

retic

ulat

e

P a

ff g

loch

idio

ides

4-

colp

orat

e su

bpro

late

19

1ndash

208

by

158

ndash17

5 1

19ndash1

21

retic

ulat

e

P

sec

urin

egoi

des

type

P

sec

urin

egoi

des

4-co

lpro

late

pr

olat

e sp

hero

idal

22

9ndash

243

by

219

ndash22

9

100

ndash11

1 re

ticul

ate

Isoc

ladu

s P

arap

hylla

nthu

s P

ery

thro

tric

hus

type

P

ery

thro

tric

hus

3-co

lpor

ate

prol

ate

to p

erpr

olat

e 15

3ndash1

93

by 9

3ndash1

00

158

ndash2

08

stria

ted

retic

ulat

e

Par

aphy

llant

hus

P v

irgat

us ty

pe

P la

ncei

foliu

s ar

eola

te

sphe

roid

al to

elli

psoi

dal

175

ndash22

5 b

y 17

5ndash1

96

ndash co

arse

ly r

etic

ulat

e

Mac

raea

P

virg

atus

type

P

sam

aren

sis

areo

late

sp

hero

idal

to e

llips

oida

l 15

3ndash1

67

by 1

50

ndash16

3 ndash

coar

sely

ret

icul

ate

P

virg

atus

ar

eola

te

sphe

roid

al to

elli

psoi

dal

167

ndash18

8 by

15

8ndash1

88

ndash re

ticul

ate

P

aff

sam

aren

sis

ar

eola

te

sphe

roid

al to

elli

psoi

dal

183

ndash19

6 by

19

2ndash

200

ndash

retic

ulat

e

Kirg

anel

ia

Ani

sone

ma

P c

astic

um ty

pe

P r

etic

ulat

us

3-sy

ncol

pora

te

obla

te s

pher

oida

l 11

1ndash1

17

by 1

17ndash

125

0

91ndash

096

re

ticul

ate

C

icca

P

aci

dus

type

P

aci

dus

3-co

lpor

ate

obla

te s

pher

oida

l 12

3ndash1

33

by 1

33

ndash15

0 0

89ndash

095

ru

gula

te r

etic

ulat

e

Phy

llant

hus

Phy

llant

hus

P a

mar

us ty

pe

P a

mar

us

3-co

lpor

ate

prol

ate

sper

oida

l to

subp

rola

te

153

ndash17

0 by

13

0ndash1

57

106

ndash13

1 bi

retic

ulat

e

Urin

aria

P

urin

aria

type

P

ben

guet

ensi

s 4-

colp

rola

te

prol

ate

200

ndash22

1 b

y 12

1ndash1

33

163

ndash17

6 bi

retic

ulat

e

P u

rinar

ia s

sp n

udic

arpu

s 4-

colp

rola

te

prol

ate

225

ndash23

8 b

y 12

9ndash1

46

163

ndash17

4 bi

retic

ulat

e

P u

rinar

ia s

sp u

rinar

ia

4-co

lpro

late

pr

olat

e 16

0ndash1

63

by 1

1ndash12

1

36ndash1

45

bire

ticul

ate

not a

ssig

ned

not a

ssig

ned

Pila

te ty

pe

Phy

llant

hus

sp A

4-

colp

orat

e su

bpro

late

14

3 b

y 11

ndash12

3 1

16ndash1

30

pila

te

Tab

le 2

P

olle

n ty

pes

of P

hilip

pine

Phy

llant

hus

spec

ies



cb

d

a

g h i

j k l

e f



cb

d

a

g h i

j k l

e f



cb

d

a

e f


cb

d

a

e f
















cb

d

a

e f



cb

d

a

g h i

j k l

e f


DISCUSSION










cb

d

a

e f



cb

d

a

g h i

j k l

e f




cb

d

a

e f

cb

d

a

e f



REFERENCES



















































Sub

genu

s S

ectio

n P

olle

n ty

pe

Taxa

A

pert

ure

Equ

ator

ial s

hape

S

ize

P b

y E

(micro

m)

PE

O

rnam

enta

tion

Erio

cocc

us

Sce

pasm

a P

bux

ifoliu

s ty

pe

P b

uxifo

lius

num

erou

s sp

hero

idal

to e

llips

oida

l 22

2ndash

224

by

244

ndash25

6

ndash re

ticul

ate

E

mbl

icas

trum

P

bux

ifoliu

s ty

pe

P la

mpr

ophy

llus

num

erou

s sp

hero

idal

to e

llips

oida

l 22

9ndash

246

by

213

ndash22

9

ndash re

ticul

ate

E

rioco

ccus

P

bux

ifoliu

s ty

pe

P k

inab

alui

cus

num

erou

s sp

hero

idal

to e

llips

oida

l 20

0ndash

204

by

200

ndash20

4

ndash re

ticul

ate

P

meg

alan

thus

nu

mer

ous

sphe

roid

al to

elli

psoi

dal

250

by

244

ndash26

1

ndash re

ticul

ate

P

min

dore

nsis

nu

mer

ous

sphe

roid

al to

elli

psoi

dal

175

ndash18

0 by

18

0 ndash

retic

ulat

e

P s

ibuy

anen

sis

num

erou

s sp

hero

idal

to e

llips

oida

l 18

0ndash

200

by

183

ndash19

6 ndash

retic

ulat

e

Gom

phid

ium

A

deno

gloc

hidi

on

P m

ariti

mus

type

P

ram

osii

3-sy

ncol

pora

te

obla

te s

pher

oida

l 15

8ndash1

67

by 1

71

092

ndash0

98

retic

ulat

e

Gom

phid

ium

P

glo

chid

ioid

es ty

pe

P g

loch

idio

ides

4-

colp

orat

e su

bpro

late

21

7ndash

221

by

175

ndash18

8 1

15ndash1

26

retic

ulat

e

P a

ff g

loch

idio

ides

4-

colp

orat

e su

bpro

late

19

1ndash

208

by

158

ndash17

5 1

19ndash1

21

retic

ulat

e

P

sec

urin

egoi

des

type

P

sec

urin

egoi

des

4-co

lpro

late

pr

olat

e sp

hero

idal

22

9ndash

243

by

219

ndash22

9

100

ndash11

1 re

ticul

ate

Isoc

ladu

s P

arap

hylla

nthu

s P

ery

thro

tric

hus

type

P

ery

thro

tric

hus

3-co

lpor

ate

prol

ate

to p

erpr

olat

e 15

3ndash1

93

by 9

3ndash1

00

158

ndash2

08

stria

ted

retic

ulat

e

Par

aphy

llant

hus

P v

irgat

us ty

pe

P la

ncei

foliu

s ar

eola

te

sphe

roid

al to

elli

psoi

dal

175

ndash22

5 b

y 17

5ndash1

96

ndash co

arse

ly r

etic

ulat

e

Mac

raea

P

virg

atus

type

P

sam

aren

sis

areo

late

sp

hero

idal

to e

llips

oida

l 15

3ndash1

67

by 1

50

ndash16

3 ndash

coar

sely

ret

icul

ate

P

virg

atus

ar

eola

te

sphe

roid

al to

elli

psoi

dal

167

ndash18

8 by

15

8ndash1

88

ndash re

ticul

ate

P

aff

sam

aren

sis

ar

eola

te

sphe

roid

al to

elli

psoi

dal

183

ndash19

6 by

19

2ndash

200

ndash

retic

ulat

e

Kirg

anel

ia

Ani

sone

ma

P c

astic

um ty

pe

P r

etic

ulat

us

3-sy

ncol

pora

te

obla

te s

pher

oida

l 11

1ndash1

17

by 1

17ndash

125

0

91ndash

096

re

ticul

ate

C

icca

P

aci

dus

type

P

aci

dus

3-co

lpor

ate

obla

te s

pher

oida

l 12

3ndash1

33

by 1

33

ndash15

0 0

89ndash

095

ru

gula

te r

etic

ulat

e

Phy

llant

hus

Phy

llant

hus

P a

mar

us ty

pe

P a

mar

us

3-co

lpor

ate

prol

ate

sper

oida

l to

subp

rola

te

153

ndash17

0 by

13

0ndash1

57

106

ndash13

1 bi

retic

ulat

e

Urin

aria

P

urin

aria

type

P

ben

guet

ensi

s 4-

colp

rola

te

prol

ate

200

ndash22

1 b

y 12

1ndash1

33

163

ndash17

6 bi

retic

ulat

e

P u

rinar

ia s

sp n

udic

arpu

s 4-

colp

rola

te

prol

ate

225

ndash23

8 b

y 12

9ndash1

46

163

ndash17

4 bi

retic

ulat

e

P u

rinar

ia s

sp u

rinar

ia

4-co

lpro

late

pr

olat

e 16

0ndash1

63

by 1

1ndash12

1

36ndash1

45

bire

ticul

ate

not a

ssig

ned

not a

ssig

ned

Pila

te ty

pe

Phy

llant

hus

sp A

4-

colp

orat

e su

bpro

late

14

3 b

y 11

ndash12

3 1

16ndash1

30

pila

te

Tab

le 2

P

olle

n ty

pes

of P

hilip

pine

Phy

llant

hus

spec

ies



cb

d

a

g h i

j k l

e f



cb

d

a

g h i

j k l

e f



cb

d

a

e f


cb

d

a

e f
















cb

d

a

e f



cb

d

a

g h i

j k l

e f


DISCUSSION










cb

d

a

e f



cb

d

a

g h i

j k l

e f




cb

d

a

e f

cb

d

a

e f



REFERENCES









































cb

d

a

g h i

j k l

e f



cb

d

a

g h i

j k l

e f



cb

d

a

e f


cb

d

a

e f
















cb

d

a

e f



cb

d

a

g h i

j k l

e f


DISCUSSION










cb

d

a

e f



cb

d

a

g h i

j k l

e f




cb

d

a

e f

cb

d

a

e f



REFERENCES









































cb

d

a

g h i

j k l

e f



cb

d

a

e f


cb

d

a

e f
















cb

d

a

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cb

d

a

g h i

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DISCUSSION










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cb

d

a

g h i

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cb

d

a

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cb

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REFERENCES









































cb

d

a

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cb

d

a

e f
















cb

d

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DISCUSSION










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cb

d

a

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cb

d

a

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cb

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REFERENCES






















































cb

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DISCUSSION










cb

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a

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cb

d

a

g h i

j k l

e f




cb

d

a

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cb

d

a

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REFERENCES









































cb

d

a

g h i

j k l

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DISCUSSION










cb

d

a

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cb

d

a

g h i

j k l

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pollen morphology of philippine species of phyllanthus

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